title: Expression and purification of hepatitis C virus protease from clinical samples.
abstract: This chapter describes the procedures for production of recombinant hepatitis C virus (HCV) NS3 protease from clinical samples, which can be used in the biochemical assays to assess the impact of different drug-resistant mutations in the NS3 protein from patients under therapy of protease inhibitors. It gives the details of expression and purification of NS3 protease using the pCold vectors that contains a promoter derived from the cold-shock genes to drive the expression of NS3 protein. This robust protocol enables a medium-throughput production of HCV NS3 proteins of all genotypes and sequences derived from patient specimen.
authors.affiliation: Janssen Infectious Diseases-Diagnostics BVBA, Johnson & Johnson Corporation, Beerse, Belgium.
authors.forename: Liesbet
authors.initials: L
authors.lastname: van der Helm
mesh_terms: D003001:Cloning, Molecular; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014169:Transformation, Bacterial; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases